CN109675103A - A kind of bone-cartilage two-way function bracket and preparation method thereof based on cell 3D printing - Google Patents

A kind of bone-cartilage two-way function bracket and preparation method thereof based on cell 3D printing Download PDF

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Publication number
CN109675103A
CN109675103A CN201811463188.8A CN201811463188A CN109675103A CN 109675103 A CN109675103 A CN 109675103A CN 201811463188 A CN201811463188 A CN 201811463188A CN 109675103 A CN109675103 A CN 109675103A
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cartilage
ink
skeletonization
bone
bracket
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郭吕华
武敬文
任文
魏娟
梁婷婷
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Stomatological Hospital of Guangzhou Medical University
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Stomatological Hospital of Guangzhou Medical University
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    • A61L27/14—Macromolecular materials
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    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
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Abstract

The invention discloses a kind of bone-cartilage two-way function bracket and preparation method thereof based on cell 3D printing, the preparation method of the bone-cartilage two-way function bracket based on cell 3D printing include the following steps: that S1. prepares skeletonization ink matrix and at cartilage ink matrix respectively；S2. mesenchymal stem cell and aurantiin are added into skeletonization ink matrix, obtains skeletonization ink；To at mesenchymal stem cell and transforming growth factor β are added in cartilage ink matrix, cartilage ink is obtained into；S3. it is printed respectively by skeletonization ink and at cartilage ink, obtains bracket；The bracket includes skeletonization side and at cartilage side；S4. by bracket after crosslinking Treatment, bone-cartilage two-way function bracket is obtained.Bone-cartilage two-way function bracket prepared by the present invention can be realized simultaneously the combining and regenerating of bone-cartilage, and realize personalized customization, be conducive to the regeneration in joint, so as to preferably repair joint injury.

Description

A kind of bone-cartilage two-way function bracket and preparation method thereof based on cell 3D printing
Technical field
The present invention relates to field of tissue engineering technology, more particularly, to a kind of two-way function of the bone-cartilage based on cell 3D printing Energy bracket and preparation method thereof.
Background technique
Bone tissue has self-regeneration and regeneration Reconstruction of The Function, and since wound, tumour, infection, aging of population are drawn Large area bone defect caused by osteoporosis risen etc., is only limited by the self-repairing capability of bone, especially joint Cartilage is unable to self-heal after damage.Current clinically common treatment method mainly has microfrature, Autologous Chondrocyte to move Plant and prosthetic replacement etc., these restorative procedures respectively have disadvantage.The rise of organizational project provides for cartilage defect repair research One new direction.
Rapid shaping technique is also known as 3D printing technique.Low temperature rapid prototyping & manufacturing technology (LDM) is based on rapid shaping skill Art principle, in conjunction with a kind of novel rapid shaping technique of phase separation method.Currently, there is researcher to be adsorbed on active growth factor LDM is printed on three-dimensional rack, the results showed that active growth factor can be sustained quickly, and bracket is enable to lose long-term inducing tissue regeneration Power.The characteristics of not needing heating using LDM shaped support process, researcher attempt bioactive molecule directly and after solution mixing to beat Print, and this will cause the burst release of bioactive molecule, and still be difficult to solve the problems, such as to discharge steadily in the long term.
It also needs to be sintered or be further processed after shaping existing bracket (such as CN105031718A), can not be implanted into Living cells, initial stage need to raise external cellular and are colonized in rack surface, have very big uncertainty, it is difficult to realize that regeneration of joints is repaired It is multiple.Also, joint injury generally will appear the combined injuries of bone-cartilage, and existing bracket is usually used in bone tissue reparation regeneration, difficult To realize the joint reparative regeneration of bone-cartilage tissue.
Therefore, it is necessary to prepare a kind of bracket for realizing bone-cartilage tissue joint reparative regeneration.
Summary of the invention
The present invention is the defect for overcoming cradling function described in the above-mentioned prior art single, provides one kind and is beaten based on cell 3D The preparation method of the bone-cartilage two-way function bracket of print.Bone-cartilage two-way function bracket can be simultaneously made from the preparation method It realizes the combining and regenerating of bone-cartilage, and personalized customization may be implemented, be conducive to the regeneration in joint, so as to more Repair joint injury well.
Another object of the present invention is to provide bone-cartilage two-way function brackets made from above-mentioned preparation method.
In order to solve the above technical problems, the technical solution adopted by the present invention is that:
A kind of preparation method of the bone-cartilage two-way function bracket based on cell 3D printing, includes the following steps:
S1. skeletonization ink matrix is prepared respectively and at cartilage ink matrix；
S2. mesenchymal stem cell and aurantiin are added into skeletonization ink matrix, obtains skeletonization ink；To at cartilage ink Mesenchymal stem cell and transforming growth factor β are added in water-based, obtains into cartilage ink；
S3. it is printed respectively by skeletonization ink and at cartilage ink, obtains bracket；The bracket includes skeletonization side and at soft Bone side；
S4. by bracket after crosslinking Treatment, bone-cartilage two-way function bracket is obtained.
Mesenchymal stem cell (BMSCs) be it is isolated from marrow, because its materials is convenient and to body injury it is small, Proliferative capacity is strong, has multi-lineage potential, and BMSCs is the building most common cell of organizational project, is widely used in tissue and lacks The reparation of damage.BMSCs has the advantage that from a wealth of sources as seed cell, is easy to draw materials；It is easy to expand, amplification in vitro energy Power is extremely strong；With unique split form, feasible cell transfection technique is implanted into foreign gene；Immunogenicity is low, can after plant people Keep good biological activity.For BMSCs as the seed cell for being applied to bone tissue engineer earliest, skeletonization effect is true It cuts, and amplification in vitro fertility is widely used by force.In addition, the potentiality of BMSCs to Chondrocyte Differentiation are definite, it is considered It is the optimal seed cell that cartilage defect repair is used in organizational project.At BMSCs Hui Zhizhi cartilage defect, embryo reproduce Bone cartilage formation process when fetal hair is educated, and the morphosis of its cartilage and normal articular cartilage are very close.
Transforming growth factor β (TGF-β) is a major class growth factor family, has and promotees embry ogenesis, tumour formation, exempts from Many biological actions such as epidemic disease reaction.BMSCs as seed cell, break up to chondrocyte phenotype it is limited by many factors, Its transforming growth factor beta is cell factor necessary to BMSCs breaks up to cartilage cell direction.
The present invention is based on cell 3D printing technique, is prepared for by preparing celliferous skeletonization ink and at cartilage ink A kind of bone-cartilage two-way function bracket.The bone-cartilage two-way function bracket has preferable intensity, and is not necessarily to high-temperature process, can To print on transforming growth factor β and living cells in bracket, conventional stent growth factor burst release and cell are avoided without legal The problem of plant.Used aurantiin is a kind of flavones monomeric substance, with chemical structure is clear, molecular weight is small, purification is convenient The features such as, have the effects that anti-oxidant, anti-cholesterol, neuroprotection, anti-inflammatory, Bone formation, induced osteogenesis, inhibit osteoclasia. Bone-cartilage two-way function bracket provided by the invention is that cell is implanted into bracket, can be realized simultaneously the joint of bone-cartilage again It is raw, and personalized customization is realized, be conducive to the regeneration in joint, so as to preferably repair joint injury.
Preferably, the concentration of the aurantiin in skeletonization ink described in step S2. is 1 × 10-4~1×10-6 mol/L.More Preferably, the concentration of the aurantiin in skeletonization ink described in step S2. is 1 × 10-5 mol/L。
Preferably, the concentration of mesenchymal stem cell described in step S2. is 3 × 106~8×106 /mL.More preferably Ground, the concentration of mesenchymal stem cell described in step S2. are 5 × 106 /mL.Preferably, between marrow described in step S2. Mesenchymal stem cells are added in the form of mesenchymal stem cell suspension.
Preferably, mesenchymal stem cell suspension described in step S2. and skeletonization ink or the volume at cartilage ink Than being 1: 3 ~ 5.
It is highly preferred that mesenchymal stem cell suspension described in step S2. and skeletonization ink or the body at cartilage ink Product is than being 1: 4.
Preferably, at the concentration 1 × 10 of the transforming growth factor β in cartilage ink described in step S2.-8 ~1×10-10 mol/L.It is highly preferred that described in step S2. at the concentration of the transforming growth factor β in cartilage ink be 1 × 10-9 mol/L。
Preferably, skeletonization ink described in step S1. includes inorganic particle and biodegradable material；Institute in step S1. Stating into cartilage ink includes biodegradable material.
Preferably, inorganic particle described in step S1. is mesoporous bioglass powder, calcium sulfate, tricalcium silicate, calcium carbonate One or more of.It is highly preferred that inorganic particle described in step S1. is mesoporous bioglass powder.
Preferably, biodegradable material described in step S1. is sodium alginate, gelatin, starch, agarose, chitosan One or more of.It is highly preferred that biodegradable material described in step S1. is sodium alginate and gelatin.
Preferably, crosslinking described in step S4. is chemical crosslinking.Preferably, the crosslinking of chemical crosslinking described in step S4. Agent is CaCl2Solution.Preferably, CaCl described in step S4.2The concentration of solution is 1% ~ 3%.It is highly preferred that institute in step S4. State CaCl2The concentration of solution is 1%.CaCl is put by the skeletonization side printed and at cartilage side2It impregnates 5 minutes, obtains in solution To bone-cartilage two-way function bracket.Chemical crosslinking increases the intensity of bone-cartilage two-way function bracket, and the bracket is not necessarily to High-temperature process.
Preferably, skeletonization ink described in step S1. includes the composition of following mass percent: mesoporous bioglass powder 5% ~ 15%, sodium alginate 5% ~ 10%, gelatin 2.5% ~ 5%, surplus is water；It at cartilage ink include following matter described in step S1. Measure the composition of percentage: sodium alginate 5% ~ 10%, gelatin 2.5% ~ 5%, surplus are water.Skeletonization ink and in cartilage ink, institute It is deionized water with water.
It is highly preferred that skeletonization ink described in step S1. includes the composition of following mass percent: mesoporous bioglass powder End 10%, sodium alginate 5%, gelatin 5%, surplus is water；It at cartilage ink include following mass percent described in step S1. Composition: sodium alginate 10%, gelatin 2.5%, surplus are water.
The present invention protects bone-cartilage two-way function bracket made from above-mentioned preparation method simultaneously.
Compared with prior art, the beneficial effects of the present invention are:
The present invention is by preparing celliferous skeletonization ink and being prepared for one kind by cell 3D printing technique at cartilage ink Bone-cartilage two-way function bracket.Cell is implanted into bracket, can be realized simultaneously the combining and regenerating of bone-cartilage, and is realized a Propertyization customization, is conducive to the regeneration in joint, so as to preferably repair joint injury.
Detailed description of the invention
Fig. 1 is the flow chart that the present invention prepares bone-cartilage two-way function bracket.
Fig. 2 is the outside drawing of bone-cartilage two-way function bracket prepared by embodiment 1.
Fig. 3 is the scanning electron microscope (SEM) photograph of bone-cartilage two-way function bracket prepared by embodiment 1.
Fig. 4 is the fluorescent staining lab diagram of bone-cartilage two-way function bracket prepared by embodiment 1.
Fig. 5 is the cell proliferation rate figure of bone-cartilage two-way function bracket prepared by embodiment 1.
Specific embodiment
The invention will be further described With reference to embodiment, but embodiments of the present invention are not limited to This.Raw material in embodiment can be by being commercially available；Unless stated otherwise, the present invention uses reagent, method and apparatus for The art conventional reagent, method and apparatus.
Embodiment 1
A kind of preparation method of bone-cartilage two-way function bracket, includes the following steps:
S1. skeletonization ink matrix and at cartilage ink matrix is configured, it is after disinfection to reserve；Wherein, skeletonization ink matrix includes as follows The composition of mass percent: mesoporous bioglass powder 10%, sodium alginate 5%, gelatin 5%, surplus are water；
Include the composition of following mass percent at cartilage ink matrix: sodium alginate 10%, gelatin 2.5%, surplus are water.
S2. mesenchymal stem cell (BMSCs) and aurantiin (Naringin) are added in skeletonization ink matrix, are obtained To skeletonization ink；In skeletonization ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of aurantiin are 1 × 10-5 mol/L。
By BMSCs and transforming growth factor β (TGF-β) (1 × 10-9Mol/L it) is added in cartilage ink matrix, obtains into Cartilage ink.At in cartilage ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of TGF-β are 1 × 10-9 mol/L。
Suspension containing mesenchymal stem cell is 1: 4 with skeletonization ink matrix or at the volume ratio of cartilage ink matrix.
S3. it is respectively put into different printing barrels by skeletonization ink and at cartilage ink respectively, adjusts corresponding printing (skeletonization side: air pressure: 0.6 Mpa jet diameters: 0.41 mm wire vent speed: 8 mm/s are at cartilage side: air pressure: 0.3 Mpa for parameter Jet diameters: 0.41 mm wire vent speed: 6 mm/s), layering printing is obtained by skeletonization side and the bracket formed at cartilage side.
S4., bracket is put into 1% CaCl2Impregnated 5 minutes in solution, obtain bone-cartilage two-way function bracket, culture to With.
Embodiment 2
A kind of preparation method of bone-cartilage two-way function bracket, includes the following steps:
S1. skeletonization ink matrix and at cartilage ink matrix is configured, it is after disinfection to reserve；Wherein, skeletonization ink matrix includes as follows The composition of mass percent: mesoporous bioglass powder 5%, sodium alginate 10%, gelatin 2.5%, surplus are water；
Include the composition of following mass percent at cartilage ink matrix: sodium alginate 10%, gelatin 5%, surplus are water.
S2. mesenchymal stem cell (BMSCs) and aurantiin (Naringin) are added in skeletonization ink matrix, are obtained To skeletonization ink；In skeletonization ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of aurantiin are 1 × 10-5 mol/L。
BMSCs and transforming growth factor β (TGF-β) are added in cartilage ink matrix, cartilage ink is obtained into.At soft In bone ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of TGF-β are 1 × 10-9 mol/L。
Suspension containing mesenchymal stem cell is 1: 5 with skeletonization ink matrix or at the volume ratio of cartilage ink matrix.
S3. it is respectively put into different printing barrels by skeletonization ink and at cartilage ink respectively, adjusts corresponding printing (skeletonization side: air pressure: 0.55 Mpa jet diameters: 0.41 mm wire vent speed: 8 mm/s are at cartilage side: air pressure: 0.25 for parameter Mpa jet diameters: 0.41 mm wire vent speed: 6 mm/s), layering printing is obtained by skeletonization side and the branch formed at cartilage side Frame.
S4., bracket is put into 3% CaCl2Impregnated 5 minutes in solution, obtain bone-cartilage two-way function bracket, culture to With.
Embodiment 3
A kind of preparation method of bone-cartilage two-way function bracket, includes the following steps:
S1. skeletonization ink matrix and at cartilage ink matrix is configured, it is after disinfection to reserve；Wherein, skeletonization ink matrix includes as follows The composition of mass percent: mesoporous bioglass powder 5%, sodium alginate 10%, gelatin 2.5%, surplus are water；
Include the composition of following mass percent at cartilage ink matrix: sodium alginate 10%, gelatin 5%, surplus are water.
S2. mesenchymal stem cell (BMSCs) and aurantiin (Naringin) are added in skeletonization ink matrix, are obtained To skeletonization ink；In skeletonization ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of aurantiin are 1 × 10-4 mol/L。
BMSCs and transforming growth factor β (TGF-β) are added in cartilage ink matrix, cartilage ink is obtained into.At soft In bone ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of TGF-β are 1 × 10-8 mol/L。
Suspension containing mesenchymal stem cell is 1: 5 with skeletonization ink matrix or at the volume ratio of cartilage ink matrix.
S3. it is respectively put into different printing barrels by skeletonization ink and at cartilage ink respectively, adjusts corresponding printing (skeletonization side: air pressure: 0.55 Mpa jet diameters: 0.41 mm wire vent speed: 8 mm/s are at cartilage side: air pressure: 0.25 for parameter Mpa jet diameters: 0.41 mm wire vent speed: 6 mm/s), layering printing is obtained by skeletonization side and the branch formed at cartilage side Frame.
S4., bracket is put into 3% CaCl2Impregnated 5 minutes in solution, obtain bone-cartilage two-way function bracket, culture to With.
Embodiment 4
A kind of preparation method of bone-cartilage two-way function bracket, includes the following steps:
S1. skeletonization ink matrix and at cartilage ink matrix is configured, it is after disinfection to reserve；Wherein, skeletonization ink matrix includes as follows The composition of mass percent: mesoporous bioglass powder 10%, sodium alginate 5%, gelatin 5%, surplus are water；
Include the composition of following mass percent at cartilage ink matrix: sodium alginate 10%, gelatin 2.5%, surplus are water.
S2. mesenchymal stem cell (BMSCs) and aurantiin (Naringin) are added in skeletonization ink matrix, are obtained To skeletonization ink；In skeletonization ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of aurantiin are 1 × 10-6 mol/L。
By BMSCs and transforming growth factor β (TGF-β) (1 × 10-9Mol/L it) is added in cartilage ink matrix, obtains into Cartilage ink.At in cartilage ink, the concentration of BMSCs is 5 × 106 / mL, the concentration of TGF-β are 1 × 10-10 mol/L。
Suspension containing mesenchymal stem cell is 1: 4 with skeletonization ink matrix or at the volume ratio of cartilage ink matrix.
S3. it is respectively put into different printing barrels by skeletonization ink and at cartilage ink respectively, adjusts corresponding printing (skeletonization side: air pressure: 0.6 Mpa jet diameters: 0.41 mm wire vent speed: 8 mm/s are at cartilage side: air pressure: 0.3 Mpa for parameter Jet diameters: 0.41 mm wire vent speed: 6 mm/s), layering printing is obtained by skeletonization side and the bracket formed at cartilage side.
S4., bracket is put into 1% CaCl2Impregnated 5 minutes in solution, obtain bone-cartilage two-way function bracket, culture to With.
The characterization and performance test of bone-cartilage two-way function bracket
1, bracket appearance
Fig. 2 is the outside drawing of bone-cartilage two-way function bracket prepared by embodiment 1.Fig. 2 shows bone-cartilage two-way function bracket Shape is good, and hole is uniform, has certain intensity and elasticity.
2, scanning electron microscope (SEM) is observed
The surface texture of bone-cartilage two-way function bracket prepared by the present invention is observed by scanning electron microscope (SEM), embodiment 1 As a result as shown in Figure 3.It skeletonization side and is slightly different at the rack surface structure of cartilage side, can be observed on skeletonization side stand surface To equally distributed mesoporous bioglass powder.Rough rack surface is conducive to cell and is colonized and adheres to, and skeletonization side is mesoporous The addition of bio-vitric is conducive to the regeneration of bone tissue.
3, fluorescent staining is tested:
Experimental procedure: being layered on 48 orifice plate bottoms (three repeating samples) for the bracket (10mm × 3mm) that the present invention prints respectively, And corresponding induced medium is added, cultivate 7 days stand-by (changing liquid in every 2 days)；With without Ca2+、Mg2+PBS, rinse 3 times, often Secondary 3 min；By specification configures corresponding staining reagent, and the staining reagent of 100 μ L is added in every hole, is protected from light and is incubated for 30 min；It goes Except staining reagent, using without Ca2+、Mg2+PBS, rinse 3 times, 3 min every time；It is thin using confocal laser scanning microscope Cytoactive.
Experimental result: Calcein-AM can pass through cell membrane, sloughs AM group by the esterase effect in living cells, generates Calcein (calcein) issue strong green fluorescence, therefore living cells can be detected green fluorescence under fluorescence microscope. Another aspect PI can enter the DNA double spiral in dead cell and being embedded in cell by impaired cell membrane to generate red Fluorescence, therefore dead cell can be detected red fluorescence.
By fluorescent staining Germicidal efficacy internal stent cell activity, Fig. 4 is the fluorescent staining of bracket prepared by embodiment 1 Lab diagram.It can be observed that the fluorescent marker of green, shows that the cell activity in bracket is good.
4, CCK-8 is tested:
Experimental procedure: the bracket (5mm × 3mm) that the present invention prints is layered on 96 orifice plate bottoms (6 repeating samples) respectively, and Corresponding 100 μ L of induced medium is added, cultivates 1,3,5,7 day stand-by (changing liquid in every two days) respectively；Former culture medium is removed, newly Fresh culture medium is mixed with CCK-8 reagent according to 10:1, and 130 μ L are added in each hole, is protected from light and is incubated for 2 h；Draw 110 μ L in each hole 96 new orifice plates are added in liquid after incubation, measure absorbance value at 450 nm of microplate reader.
Experimental result: contain WST-8, the chemical name of WST-8 in CCK-8 reagent are as follows: 2- (2- methoxyl group -4- nitrobenzene Base) -3- (4- nitrobenzophenone) -5- (2,4- disulfonic acid benzene) -2H- tetrazolium monosodium salt.WST-8 is in electron carrier 1- methoxyl group -5- It is reduced to have the yellow first a ceremonial jade-ladle, used in libation of high water soluble to produce by the dehydrogenase in cell under the action of toluphenazine dimethyl suflfate Object.The quantity of the first a ceremonial jade-ladle, used in libation object of generation is directly proportional to the quantity of living cells.
By the cell proliferation rate in CCK-8 measuring bracket, Fig. 5 is that the cell of bracket prepared by embodiment 1 increases Grow rate diagram.After 7 days, skeletonization side and the CCK-8 numerical value no significant difference at cartilage side stand show bracket inner cell number Amount does not occur had significant proliferation or apoptosis.
It follows that bone-cartilage two-way function bracket prepared by the present invention is cell can be implanted into bracket, while it is real The combining and regenerating of existing bone-cartilage, and realize personalized customization.Bone-cartilage two-way function bracket is conducive to the tissue in joint again It is raw, so as to preferably repair joint injury.
Obviously, the above embodiment of the present invention be only to clearly illustrate example of the present invention, and not be pair The restriction of embodiments of the present invention.For those of ordinary skill in the art, may be used also on the basis of the above description To make other variations or changes in different ways.There is no necessity and possibility to exhaust all the enbodiments.It is all this Made any modifications, equivalent replacements, and improvements etc., should be included in the claims in the present invention within the spirit and principle of invention Protection scope within.

Claims (10)

1. a kind of preparation method of the bone-cartilage two-way function bracket based on cell 3D printing, which is characterized in that including walking as follows It is rapid:
S1. skeletonization ink matrix is prepared respectively and at cartilage ink matrix；
S2. mesenchymal stem cell and aurantiin are added into skeletonization ink matrix, obtains skeletonization ink；To at cartilage ink Mesenchymal stem cell and transforming growth factor β are added in water-based, obtains into cartilage ink；
S3. it is printed respectively by skeletonization ink and at cartilage ink, obtains bracket；The bracket includes skeletonization side and at soft Bone side；
S4. by bracket after crosslinking Treatment, bone-cartilage two-way function bracket is obtained.
2. preparation method according to claim 1, which is characterized in that the aurantiin in skeletonization ink described in step S2. Concentration be 1 × 10-4~1×10-6 mol/L。
3. preparation method according to claim 2, which is characterized in that the aurantiin in skeletonization ink described in step S2. Concentration be 1 × 10-5 mol/L。
4. preparation method according to claim 1, which is characterized in that mesenchymal stem cell described in step S2. Concentration is 3 × 106~8×106 /mL。
5. the preparation method according to claim 4, which is characterized in that mesenchymal stem cell described in step S2. Concentration is 5 × 106/mL。
6. preparation method according to claim 1, which is characterized in that at the conversion in cartilage ink described in step S2. The concentration of grouth factor beta is 1 × 10-8 ~1×10-10 mol/L。
7. preparation method according to claim 1, which is characterized in that at the conversion in cartilage ink described in step S2. The concentration of grouth factor beta is 1 × 10-9 mol/L。
8. preparation method according to claim 1, which is characterized in that skeletonization ink described in step S1. includes inorganic powder Body and biodegradable material；Include biodegradable material at cartilage ink described in step S1..
9. preparation method according to claim 8, which is characterized in that inorganic particle described in step S1. is mesoporous biological One or more of glass powder, calcium sulfate, tricalcium silicate, calcium carbonate.
10. bone-cartilage two-way function bracket made from the described in any item preparation methods of claim 1 ~ 9.
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Application publication date: 20190426